fig1

Figure 1. Optimization of the PROSPR protocol for EV enrichment from blood samples. (A) Average EV particle concentration (particles/mL) measured by dFC across varying starting volumes of blood (50, 100 and 200 μL); (B) Average EV protein concentration (mg/mL) quantified by BCA assay for the same blood volumes; (C) Correlation between average EV particle concentration and average protein concentration for each initial blood volume condition. Pearson correlation analysis was applied to evaluate significance; (D) EV size distribution (nm) and particle concentration (particles/mL) obtained by TRPS from PROSPR-enriched EVs from blood. Discontinuous lines indicate SEM. Statistical significance was assessed using one-way ANOVA (^*P < 0.05, ^**P < 0.01, ^***P < 0.001). EV: Extracellular vesicle; dFC: dedicated flow cytometry; BCA: bicinchoninic acid; TRPS: tunable resistive pulse sensing; PROSPR: PRotein Organic Solvent PRecipitation; SEM: standard error of the mean; ANOVA: analysis of variance.